Qi Jieying, Xue Qi, Kuang Liuyan, Xie Lifen, Luo Ren, Nie Xiaoli
School of Traditional Chinese Medicine, Southern Medical University, Guangzhou, China.
Hospital of Integrated Traditional Chinese Medicine and Western Medicine, Southern Medical University, Guangzhou, China.
Transl Androl Urol. 2020 Aug;9(4):1712-1724. doi: 10.21037/tau-20-1129.
To study the protective effect of berberine (BBR) on cisplatin-induced acute kidney injury (AKI) and its effect on mitophagy.
(I) Male C57BL/6 mice aged 6-8 weeks were randomly divided into control group (saline), cisplatin group (cisplatin), and cisplatin + BBR (5, 10 mg/kg) groups. In the cisplatin group and BBR groups, mice were injected intraperitoneally with 15 mg/kg of cisplatin. Mice in BBR groups were given BBR at 72, 48, 24, 0.5 h before and 24, 48 h after cisplatin injection. Mice were sacrificed 72 h after cisplatin injection, and blood were collected for detecting serum creatinine (SCr) and blood urea nitrogen (BUN) levels. Kidneys were collected for detecting protein expression levels of Kidney injury molecule 1 (KIM-1), LC3 II/LC3 I, p62, PINK 1, Parkin in the renal tissue by Western blotting. The pathological changes in renal tissues were observed using periodic acid-Schiff (PAS) staining. (II) Renal tubular epithelial cells (RTECs) were pretreated with different concentrations (1, 2, and 4 µM) of BBR, and then incubated with cisplatin. Changes in autophagy proteins LC3 II/LC3 I, p62, PINK 1, and Parkin were detected by Western blotting, and changes in cellular reactive oxygen species (ROS) and mitochondrial membrane potential (MMP) were detected by flow cytometry.
(I) Mice treated with BBR at dosage of 5 and 10 mg/kg for 6 days showed significant reduction in SCr and BUN compared to that in mice treated with cisplatin. PAS staining and immunohistochemistry showed that BBR ameliorated cisplatin-induced nephrotoxicity and reduced cisplatin-induced increase in protein expression levels of KIM-1. Compared to cisplatin-treated mice, the mice treated with BBR showed increased LC3 II/LC3 I, PINK 1, and Parkin, and decreased p62 protein expression. (II) Compared to cisplatin-incubated RTECs, cells pretreated with BBR for 24 h exhibited increased protein expressions of LC3 II/LC3 I, PINK1, and Parkin and decreased protein expression of p62. BBR reversed cellular ROS and cell MMP level induced by cisplatin.
BBR plays a protective role in cisplatin-induced AKI by up-regulating mitophagy in RTECs.
研究黄连素（BBR）对顺铂诱导的急性肾损伤（AKI）的保护作用及其对线粒体自噬的影响。
（I）将6 - 8周龄的雄性C57BL/6小鼠随机分为对照组（生理盐水）、顺铂组（顺铂）和顺铂+BBR（5、10 mg/kg）组。在顺铂组和BBR组中，小鼠腹腔注射15 mg/kg顺铂。BBR组小鼠在顺铂注射前72、48、24、0.5小时及注射后24、48小时给予BBR。顺铂注射72小时后处死小鼠，采集血液检测血清肌酐（SCr）和血尿素氮（BUN）水平。采集肾脏，通过蛋白质免疫印迹法检测肾组织中肾损伤分子1（KIM-1）、微管相关蛋白1轻链3 II/微管相关蛋白1轻链3 I（LC3 II/LC3 I）、p62、磷酸化的PTEN诱导激酶1（PINK 1）、帕金蛋白（Parkin）的蛋白表达水平。采用高碘酸-希夫（PAS）染色观察肾组织的病理变化。（II）用不同浓度（1、2和4 μM）的BBR预处理肾小管上皮细胞（RTECs），然后与顺铂共同孵育。通过蛋白质免疫印迹法检测自噬蛋白LC3 II/LC3 I、p62、PINK 1和Parkin的变化，通过流式细胞术检测细胞活性氧（ROS）和线粒体膜电位（MMP）的变化。
（I）与顺铂处理的小鼠相比，用5和10 mg/kg剂量的BBR处理6天的小鼠SCr和BUN显著降低。PAS染色和免疫组织化学显示，BBR改善了顺铂诱导的肾毒性，并降低了顺铂诱导的KIM-1蛋白表达水平的升高。与顺铂处理的小鼠相比，用BBR处理的小鼠LC3 II/LC3 I、PINK 1和Parkin增加，p62蛋白表达降低。（II）与顺铂孵育的RTECs相比，用BBR预处理24小时的细胞LC3 II/LC3 I、PINK1和Parkin的蛋白表达增加，p62蛋白表达降低。BBR逆转了顺铂诱导的细胞ROS和细胞MMP水平。
BBR通过上调RTECs中的线粒体自噬对顺铂诱导的AKI发挥保护作用。
